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pUC18Z_ePAG2

Cloning vector harboring counterselection marker Escherichia coli phenylalanyl-tRNA synthetase a-subunit (ePheS).

Catalog number RDB17948
Resource name pUC18Z_ePAG2
Clone info. Cloning vector harboring counterselection marker Escherichia coli phenylalanyl-tRNA synthetase a-subunit (ePheS) when 4-chloro-phenylalanine (4CP) is present. Zeocin resistance. See Fig. S2 of Miyazaki, K. Biotechniques, 58 (2): 86-88, 2015 for constructs.
Causing host cell death in the presence of 1 to 10% (w/v) of 4-chlorophenylalanine.
Vector backbone pUC18 (plasmid)
Selectable markers Ampicillin
Gene/insert name Escherichia coli pheS genomic DNA Streptoalloteichus hindustanus zeocin resistance gene genomic DNA
Depositor|Developer Miyazaki, Kentaro |
 
Other clones in our bank Escherichia coli (KEGG orthology K01889) |

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
Terms and conditions set forth by the DEPOSITOR In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature(s) designated by the DEPOSITOR is requested. (Miyazaki K. et al. Biotechniques, 58 (2): 86-88, 2015)
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RDB17948 pUC18Z_ePAG2 DNA solution

Ordering Information [in Japanese] [in English]
Please review the QC test results indicated by check icon below as well as clone information before placing your order.

References and tips

Electronic file

Electronic file Nucleotide sequence provided by the depositor RDB17948zzk01.seq

Original reference

original Miyazaki, K., Molecular engineering of a PheS counterselection marker for improved operating efficiency in Escherichia coli. Biotechniques 58 (2): 86-88 (2015). PMID 25652032.

Further references such as user reports and related articles (go to bottom)

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Sequence information

RIKEN BRC has sequenced portions of this material for quality test.
Please review the QC test results indicated by check icon as well as clone information before placing your order.

Test sheet RDB17948_B0Bcp1-2.pdf check

Nucleotide sequence of a portion of this resource (if available).

Primer: ZeoR-C_F
Region: insert mid (ZeoR 3', pheS 5')
Sequence file: RDB17948_B0Bca.seq check
>D06895A6_B0Bc_2_ZeoR-C_F_D03_12_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NGNNNNNNGC CGNNNNNNGC GTGCNCTTCG TGGCCGANGA
   61 NCAGGACTAA AGGAGGTAAA ACACATGTCA CATCTCGCAG AACTGGTTGC CAGTGCGAAG
  121 GCGGCCATTA GCCAGGCGTC AGATGTTGCC GCGTTAGATA ATGTGCGCGT CGAATATTTG
  181 GGTAAAAAAG GGCACTTAAC CCTTCAGATG ACGACCCTGC GTGAGCTGCC GCCAGAAGAG
  241 CGTCCGGCAG CTGGTGCGGT TATCAACGAA GCGAAAGAGC AGGTTCAGCA GGCGCTGAAT
  301 GCGCGTAAAG CGGAACTGGA AAGCGCTGCA CTGAATGCGC GTCTGGCGGC GGAAACGATT
  361 GATGTCTCTC TGCCAGGTCG TCGCATTGAA AACGGCGGTC TGCATCCGGT TACCCGTACC
  421 ATCGACCGTA TCGAAAGTTT CTTCGGTGAG CTTGGCTTTA CCGTGGCAAC CGGGCCGGAA
  481 ATCGAAGACG ATTATCATAA CTTCGATGCT CTGAACATTC CTGGTCACCA CCCGGCGCGC
  541 GCTGACCACG ACACTTTCTG GTTTGACACT ACCCGCCTGC TGCGTACCCA GACCTCTGGC
  601 GTACAGATCC GCACCATGAA AGCCCAGCAG CCACCGATTC GTATCATCGC GCCTGGCCGT
  661 GTTTATCGTA ACGACTACGA CCAGACTCAC ACGCCGATGT TCCATCAGAT GGAAGGTCTG
  721 ATTGTTGATA CCNACATCAG CTTTACCAAC CTGAAAGGNA CGCTGCACGA CTTCCTGCGT
  781 AACTTCTTTG ANGAAGANTT GCAGANTCGC TTCCGTCCTT CCTACTTCNC GTTTGCTGAN
  841 CCTTCTGCAG AAGTGGANGT CNTGGGGTAA AAACGGTAAA TGGCTGGAAN TGCTGGGCTG
  901 CGGGNNGGNG NATNCNNAAC GNGTTGCGNN NCGTNNNCNT CNANCCNGNA NNTTTACTCT
  961 GGNTTCGGNT NCNGGANNGG GANGGANCGN CNGANTATGN NGCGNNNNNG CGNNNNNNAN
 1021 NTGNGNNCNT TCNNNNNAAA CNNNNNNNNN TNNNNNNNNA GNTNNANNNN NNNNCNNGNN
1081 NNNNNNNN
//
Primer: Zeo-M_R
Region: insert 5' (ZeoR 5')
Sequence file: RDB17948_B0Bcb.seq check
>D06895A6_B0Bc_2_Zeo-M_R_F03_18_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNGCGNNNCC NCNCCCNGGC NNGGGTGTTG TCCGGCACCA
   61 CCTGGTCCTG GACCGCGCTG ATGAACAGGG TCACGTCGTC CCGGACCACA CCGGCGAAGT
  121 CGTCCTCCAC GAAGTCCCGA GAGAACCCGA GCCGGTCGGT CCAGAACTCG ACCGCTCCGG
  181 CGACGTCGCG CGCGGTGAGC ACCGGAACGG CACTGGTCAA CTTGGCCATA CTCTTCCTTT
  241 TTCAATATTA TTGAAGCATT TATCAGGGTT ATTGTCTCAT GAGCGGATAC ATATTTGAAT
  301 GTATTTAGAA AAATAAACAA ATAGGGGTTC CGCGCACATT TCCCCGAAAA GTGCCACCTG
  361 ACGTCTAAGA AACCATTATT ATCATGACAT TAACCTATAA AAATAGGCGT ATCACGAGGC
  421 CCTTTCGTCT CGCGCGTTTC GGTGATGACG GTGAAAACCT CTGACACATG CAGCTCCCGG
  481 AGACGGTCAC AGCTTGTCTG TAAGCGGATG CCGGGAGCAG ACAAGCCCGT CAGGGCGCGT
  541 CAGCGGGTGT TGGCGGGTGT CGGGGCTGGC TTAACTATGC GGCATCAGAG CAGATTGTAC
  601 TGAGAGTGCA CCATGTGCGG TGTGAAATAC CGCACAGATG CGTAAGGAGA AAATACCGCA
  661 TCAGGCGCCA TTCGCCATTC AGGCTGCGCA ACTGTTGGGA AGGGCGATCG GTGCGGGCCT
  721 CTTCGCTATT ACGCCAGCTG GCGAAAGGGG GANGTGCTGC AAGGCGATTA AGTTGGGTAA
  781 CGCCAGGGTT TTCCCAGTCA CGACGTTGTA AAACGACGGC CAGTGCCNAG CTTGCNTGNC
  841 TGCANGNCGA NTCTAGAGGN NCCCCNGNTA NCGANCTCGA ATTCNTAATC NTGGNCNTAT
  901 GTGTTTCCTG NGTGAAATTG NNATCCGCTC NNNATTCCNC NCNNNNTACN ANCCNNNNNN
  961 NTAAAGNGNA AANNNTGGGN NGNNTNNNGA NNGANCTNNC TCNNNNTNNN NNGNNNNNNN
1021 NNN
//
Primer: ePAG_R1
Region: insert 3' (pheS 3')
Sequence file: RDB17948_B0Bcc.seq check
>D06895A6_B0Bc_2_ePAG_R1_E03_15_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NTNNCCTAGN TCCTTTTAAA TTAAAAATGA AGTTTTAAAT
   61 CAATCTAAAG TATATATGAG TAAACTTGGT CTGACAGGAA TTATTATTTA AACTGTTTGA
  121 GGAAACGCAG ATCGTTTTCG AAGAATGAAC GCAGGTCGGT GACGCCGTAA CGCAACATAG
  181 TCAGACGCTC CATCCCCATC CCGAACCCGA AACCAGAGTA AACTTCCGGG TCGATGCCAA
  241 CGTTACGCAA CACGTTCGGA TGCACCATCC CGCAGCCCAG CACTTCCAGC CATTTACCGT
  301 TTTTACCCAT GACGTCCACT TCTGCAGAAG GTTCAGCAAA CGGGAAGTAG GAAGGACGGA
  361 AGCGAATCTG CAAATCTTCC TCAAAGAAGT TACGCAGGAA GTCGTGCAGC GTGCCTTTCA
  421 GGTTGGTAAA GCTGATGTTG GTATCAACAA TCAGACCTTC CATCTGATGG AACATCGGCG
  481 TGTGAGTCTG GTCGTAGTCG TTACGATAAA CACGGCCAGG CGCGATGATA CGAATCGGTG
  541 GCTGCTGGGC TTTCATGGTG CGGATCTGTA CGCCAGAGGT CTGGGTACGC AGCAGGCGGG
  601 TAGTGTCAAA CCAGAAAGTG TCGTGGTCAG CGCGCGCCGG GTGGTGACCA GGAATGTTCA
  661 GAGCATCGAA GTTATGATAA TCGTCTTCGA TTTCCGGCCC GGTTGCCACG GTAAAGCCNA
  721 GCTCACCGAA GAAACTTTCG ATACGGTCGA TGGTACGGGT AACCGGANGC AGACCGCCGT
  781 TTTCAATGCG ACGACCTGGC AGANAGACAT CNATCGTTTC CGCCGCCAGA NGCGCATTCA
  841 GTGCAGCGCT TTCCAGTTCC GCTTTACGCG CATTCNGCGN CNGNTGAACC NGCTCTTTNG
  901 CTNCNTTGAN AANCGCACCA GCTGCNNGAN GCTCNNCTGG NGGNNGCTCA CGNNNNNNGN
961 CATNNGNNNN NNNNAGNGNC NNTTTTTANC NNNNANTNNA NNNNCANANN NNNNNNNN
//
Primer: SV40pA3'Element Reverse
Region: ColE1 ori, MCS
Sequence file: RDB17948_B0Bcd.seq check
>17948_17948_B0Bc_1_SV40pA3'elementReverse_H01_08_ABI08.ab1
    1 NNNNNNNNNN GNNNNNNNNN NNNNNNNNNN NNNNNNCNNN NNNCGCGGCC TTTNNNNNNT
   61 TCCTGGCCTT TTGCTGGCCT TTTGCTCACA TGTTCTTTCC TGCGTTATCC CCTGATTCTG
  121 TGGATAACCG TATTACCGCC TTTGAGTGAG CTGATACCGC TCGCCGCAGC CGAACGACCG
  181 AGCGCAGCGA GTCAGTGAGC GAGGAAGCGG AAGAGCGCCC AATACGCAAA CCGCCTCTCC
  241 CCGCGCGTTG GCCGATTCAT TAATGCAGCT GGCACGACAG GTTTCCCGAC TGGAAAGCGG
  301 GCAGTGAGCG CAACGCAATT AATGTGAGTT AGCTCACTCA TTAGGCACCC CAGGCTTTAC
  361 ACTTTATGCT TCCGGCTCGT ATGTTGTGTG GAATTGTGAG CGGATAACAA TTTCACACAG
  421 GAAACACATA TGACCATGAT TACGAATTCG AGCTCGGTAC CCGGGGATCC TCTAGAGTCG
  481 ACCTGCAGGC ATGCAAGCTT GGCACTGGCC GTCGTTTTAC AACGTCGTGA CTGGGAAAAC
  541 NNNNNNNNNA CCCAACTTAA TCGCCTTGCA NCACATCCCC CTTTCGCCAG CTGGCGNNAA
  601 TANCGAAGAN GGNNNNNNCC NATCGCCCNT TCCNNNANAG NTNGCNCNNC CNGAATGGGN
  661 NANNGGGCGC CTGATGCNNN NNTTTTTCNC CTTTNNNCNN CTGNGNNGNA TTNNANACNN
721 CNNNNNGNNG NNCNNNNNNN ANNNNCTGCN NNNNNANGNC NNNNNNNNNN
//

Please visit Sequencing and PCR primers for primer information.


References

Original, user report and related articles

original Miyazaki, K., Molecular engineering of a PheS counterselection marker for improved operating efficiency in Escherichia coli. Biotechniques 58 (2): 86-88 (2015). PMID 25652032.

2021.01.13

GNP_filter3_RDBDEP_html_201222.pl